Circumventing the Crabtree Effect: A method to induce lactate consumption and increase oxidative phosphorylation in cell culture. (October 2016)

• Record Type: Journal Article
• Authors: Mot, Alexandra I.; Liddell, Jeffrey R.; White, Anthony R.; Crouch, Peter J.
• Is Part Of: International journal of biochemistry & cell biology. Volume 79(2016:Oct.)
• Page Start: 128
• Page End: 138
• Publication Date: 2016-10
• Subjects: AMPK α5′-adenosine monophosphate-activated protein kinase alpha -- ATP5A ATP synthase subunit alpha -- BCA bicinchoninic acid -- FCCP carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone -- COX5B cytochrome c oxidase subunit 5B -- 2-DG 2-deoxyglucose -- GAPDH glyceraldehyde-3-phosphate dehydrogenase -- OXPHOS oxidative phosphorylation -- PCNA proliferating cell nuclear antigen -- pMEFs primary mouse embryonic fibroblasts -- SDHB succinate dehydrogenase subunit B -- SOD1 superoxide dismutase 1 -- SOD2 superoxide dismutase 2 -- TOM20 mitochondrial import receptor subunit TOM20 homolog; Cell culture -- Mitochondria -- Lactate -- Oxidative phosphorylation (OXPHOS) -- Electron transport chain (ETC); Biochemistry -- Periodicals; Cytology -- Periodicals; Biochemistry -- Periodicals; Cell Biology -- Periodicals; Biochimie -- Périodiques; Cytologie -- Périodiques; Biochimie; Cytologie; Biochemistry; Cytology; Ressource Internet (Descripteur de forme); Périodique électronique (Descripteur de forme); Periodicals; 572.05
• DOI: 10.1016/j.biocel.2016.08.029
• Languages: English
• ISSNs: 1357-2725

Abstract

Abstract: Most cells grown in glucose-containing medium generate almost all their ATP via glycolysis despite abundant oxygen supply and functional mitochondria, a phenomenon known as the Crabtree effect. By contrast, most cells within the body rely on mitochondrial oxidative phosphorylation (OXPHOS) to generate the bulk of their energy supply. Thus, when utilising the accessibility of cell culture to elucidate fundamental elements of mitochondria in health and disease, it is advantageous to adopt culture conditions under which the cells have greater reliance upon OXPHOS for the supply of their energy needs. Substituting galactose for glucose in the culture medium can provide these conditions, but additional benefit can be gained from alternate in vitro models. Herein we describe culture conditions in which complete autonomous depletion of medium glucose induces a lactate-consuming phase marked by increased MitoTracker Deep Red staining intensity, increased expression of Kreb's cycle proteins, increased expression of electron transport chain subunits, and increased sensitivity to the OXPHOS inhibitor rotenone. We propose these culture conditions represent an alternate accessible model for the in vitro study of cellular processes and diseases involving the mitochondrion without limitations incurred via the Crabtree effect.